Determination of Physiological and Proteomic Responses of Metal Based Nanoparticle Treatments on Tomato Plants
Project no. 21028 · Status: Kapanmış · 2019
Summary
<p>Metal-based nanoparticle applications have an important role in plant growth and development with their widespread use. Iron and silver nanoparticles come forward because of their specific properties. In this thesis study, iron and silver nanoparticles were produced by green synthesis method. Then, the physiological and proteomic responses of the applied nanoparticles on the tomato plant were investigated. Synthesized nanoparticles were characterized by using UV-VIS spectrophotometer, SEM, EDS, FTIR, Zeta size and Zeta potential. Results showed that size silver nanoparticle were 33 nm and the size of iron nanoparticles were 34 nm. Structural conformation of nanoparticles was performed by UV-VIS, FTIR, EDS and Zeta analyses. After foliar application of iron and silver nanoparticles onto plants, macroscopic, biochemical/molecular, and proteomic observations were done, and their effects were investigated on plants. Within the scope of macroscopic observations, the iron nanoparticles showed positive effects on plant growth and development in 250 mg L-1 dose application, while 25 mg L-1 and 50 mg L-1 silver showed positive effects on root-stem lengths. The nanoparticle applications did not cause any change in H2O2 and MDA content, photosynthetic yield analysis and photosynthetic pigment analyzes made within the scope of biochemical/molecular observations. However, protein content, CAT and total antioxidant activity levels decreased in all doses of nanoparticle applications. In addition, proline and SOD contents increased in all nanoparticle doses applied. This situation revealed that nanoparticle applications can be tolerated by the plant, but at the proteomic level, the plants are exposed to oxidative stress. In the proteomic analysis, plant proteins were first separated by SDS-PAGE analysis. In the obtained gel images, there were variations on a total of 8 bands, which were at around 75, 68, 57, 43, 39, 22, 17 and 15 kDa in all nanoparticle applications. The protein band determined at a molecular weight of around 39 kDa was completely lost in silver nanoparticle applications. Fe nanoparticle applications formed a new band with a molecular weight of around 22 kDa. In other bands, decreases in band intensities were observed. Finally, in order to analyze whether this change is caused by metal binding proteins, the proteins of determined molecular weight were selected from the tomato proteome and analyzed bioinformatically. While 3373 proteins were demonstrated on the bands with <em>in silico</em> sequence analysis, a total of 114 of them were determined to bind metals. In three-dimensional structural modeling analyses, the structures of 82 proteins were verified using Phyre2, ProSA and MIB tools. The identified proteins were analyzed using CELLO and InterPro tools for their localization, biological processes involved, molecular functions, and cellular components that they participate in. Results confirm that plants cause changes in oxidative stress-related proteins at the proteomic level.</p>